Methods of use for inhibiting bone loss and lowering serum cholesterol

ABSTRACT

A method of inhibiting bone loss or resorption, or lowering serum cholesterol, comprising administering to a human in need thereof a compound having the formula ##STR1## or a pharmaceutically acceptable salt or solvate thereof, in a low dosage amount. Also encompased by the invention is a a pharmaceutical formulation in unit dosage form comprising, per unit dosage, a low dosage amount.

FIELD OF THE INVENTION

This invention relates to methods for inhibiting bone loss and loweringserum cholesterol using low dosage amounts of particular2-phenyl-3-aroylbenzothiophenes.

BACKGROUND OF THE INVENTION I. Bone Loss

The current major diseases or conditions of bone which are of publicconcern include post-menopausal osteoporosis, ovariectomy patients,senile osteoporosis, patients undergoing long-term treatment ofcorticosteroids, side effects from glucocorticoid or steroid treatment,patients suffering from Cushings's syndrome, gonadal dysgensis,periarticular erosions in rheumatoid arthritis, osteoarthritis, Paget'sdisease, osteohalisteresis, osteomalacia, hypercalcemia of malignancy,osteopenia due to bone metastases, periodontal disease, andhyperparathyroidism. All of these conditions are characterized by boneloss, resulting from an imbalance between the degradation of bone (boneresorption) and the formation of new healthy bone. This turnover of bonecontinues normally throughout life and is the mechanism by which boneregenerates. However, the conditions stated above will tip the balancetowards bone loss such that the amount of bone resorbed is inadequatelyreplaced with new bone, resulting in net bone loss.

One of the most common bone disorders is post-menopausal osteoporosiswhich affects an estimated 20 to 25 million women in the United Statesalone. Women after menopause experience an increase in the rate of boneturnover with resulting net loss of bone, as circulating estrogen levelsdecrease. The rate of bone turnover differs between bones and is highestin sites enriched with trabecular bone, such as the vertebrae and thefemoral head. The potential for bone loss at these sites immediatelyfollowing menopause is 4-5% per year. The resulting decrease in bonemass and enlargement of bone spaces leads to increased fracture risk, asthe mechanical integrity of bone deteriorates rapidly.

At present, there are 20 million people with detectable vertebralfractures due to osteoporosis and 250,000 hip fractures per yearattributable to osteoporosis in the U.S. The latter case is associatedwith a 12% mortality rate within the first two years and 30% of thepatients will require nursing home care after the fracture. Therefore,bone disorders are characterized by a noticeable mortality rate, aconsiderable decrease in the survivor's quality of life, and asignificant financial burden to families.

Essentially all of the conditions listed above would benefit fromtreatment with agents which inhibit bone resorption. Bone resorptionproceeds by the activity of specialized cells called osteoclasts.Osteoclasts are unique in their ability to resorb both thehydroxyapatite mineral and organic matrix of bone. They are identicalwith the cartilage resorbing cells, previously termed chondroclasts. Itis for this reason that potent inhibitors of osteoclastic boneresorption will also inhibit the cell-mediated degradation of cartilageobserved in rheumatoid arthritis and osteoarthritis.

Therapeutic treatments to impede net bone loss include the use ofestrogens. Estrogens have been shown clearly to arrest the bone lossobserved after menopause and limit the progression of osteoporosis;butpatient compliance has been poor because of estrogen side-effects. Theseside effects include resumption of menses, mastodynia, increase in therisk of uterine cancer, and possibly an increase in the risk of breastcancer.

Alternatively, calcitonin has been used to treat osteoporotic patients.Salmon calcitonin has been shown to directly inhibit the resorptionactivity of mammalian osteoclasts and is widely prescribed in Italy andJapan. However, calcitonins are prohibitively expensive to many andappear to be short-lived in efficacy. That is, osteoclasts are able to"escape" calcitonin inhibition of resorption by down-regulatingcalcitonin receptors. Therefore, recent clinical data suggest thatchronic treatment with calcitonin may not have long term effectivenessin arresting the post-menopausal loss of bone.

II. Serum Cholesterol

All mammalian cells require cholesterol as a structural component offtheir cell membranes and for non-sterol end products. Cholesterol isalso required for steroid hormone synthesis. The very property, however,that makes cholesterol useful in the cell membranes, its insolubility inwater, also makes it potentially lethal. When cholesterol accumulates inthe wrong place, for example within the wall of an artery, it cannot bereadily mobilized and its presence leads to the development of anatherosclerotic plaques. Elevated concentrations of serum cholesterolassociated with low density lipoproteins have been demonstrated to be amajor contributing factor in the development and progression ofatherosclerosis.

In mammals, serum lipoprotein is composed of cholesterol together withcholesterol esters, triglycerides, phospholipids and apoproteins. Serumor plasma lipoprotein is comprised of several fractions. The majorfractions or classes of plasma lipoproteins are very low densitylipoprotein (VLDL), low density lipoprotein (LDL), intermediate densitylipoprotein (IDL), and high density lipoprotein (HDL). These classesdiffer from one another in size, density and in the relative proportionsof triglycerides and cholesteryl esters in the core, and in the natureof the apoproteins on the surface.

In mammals, serum cholesterol is derived from exogenous dietary sourcesas well as through endogenous synthesis. Endogenous synthesis ofcholesterol involves a complex set of enzyme-catalyzed reactions andregulatory mechanisms generally termed the mevalonate pathway. Cellsface a complex problem in regulating mevalonate synthesis becausecholesterol, the bulk end product of mevalonate metabolism, is derivedfrom plasma low density lipoprotein which enters the cell byreceptor-mediated endocytosis, as well as from synthesis within thecell. Each cell must balance these external and internal sources so asto sustain mevalonate synthesis while avoiding sterol over accumulation.This balance is achieved through feedback regulation of at least twosequential enzymes in mevalonate synthesis, 3-hydroxy-3-methylglutarylcoenzyme A (HMG-CoA) synthase and HMG-CoA reductase and also of LDLreceptors. In the absence of LDL, mammalian cells maintain highactivities of the two enzymes, thereby synthesizing mevalonate forproduct:ion of cholesterol as well as the non-sterol products. When LDLis present, from exogenous sources, HMG-CoA synthase and reductaseactivity is repressed and the cells produce smaller amounts ofmevalonate for the non-sterol end products.

Abundant evidence indicates that treatment of hyperlipoproteinemia willdiminish or prevent atherosclerotic complications. In addition to a dietthat maintains a normal body weight and minimizes concentrations oflipids in plasma, therapeutic Strategies include elimination of factorsthat exacerbate hyperlipoproteinemia and the administration oftherapeutic agents that lower plasma concentrations of lipoproteins,either by diminishing the production of lipoproteins or by enhancing theefficiency of their removal from plasma.

The most promising class of drugs currently available for the treatmentof hypercholesterolemia act by inhibiting HMG-CoA reductase, the ratelimiting enzyme of endogenous cholesterol synthesis. Drugs of this classcompetitively inhibit the activity of the enzyme. Eventually, thislowers the endogenous synthesis of cholesterol and, by normalhomeostatic mechanisms, plasma cholesterol is taken up by LDL receptorsto restore the intracellular cholesterol balance.

Relative to other cells in the body, liver cells play a critical role inmaintaining serum cholesterol homeostasis by both releasing precursorsof LDL and through receptor mediated LDL uptake from the serum. In bothman and animal models an inverse correlation appears to exist betweenliver LDL receptors and LDL-associated serum cholesterol levels. Ingeneral, higher hepatocyte receptor numbers result in lowerLDL-associated serum cholesterol levels. Cholesterol released intohepatocytes can be stored as cholesterol esters, converted into bileacids and released into the bile duct, or enter into an oxycholesterolpool. It ms this oxycholesterol pool that is believed to be involved inend product repression of both the genes of the LDL receptor and enzymesinvolved in the cholesterol synthetic pathway.

Transcription of the LDL receptor gene is known to be repressed whencells have an excess supply of cholesterol, probably in the form ofoxycholesterol. A DNA sequence in the LDL receptor promoter region,known as the sterol response element, appears to confer this sterol endproduct repression. This element has been extensively studied (Brown,Goldstein and Russell, U.S. Pat. Nos. 4,745,060 and 4,935,363) andappears to consist of a 16 base pair sequence that occurs 5' of the LDLreceptor coding region. The sterol response element can be inserted intogenes that normally do not respond to cholesterol, conferring sterol endproduct repression on the chimeric gene. The exact mechanism of thisrepression is not understood. There is, however, abundant evidence thatpolar intermediates in cholesterol biosynthesis and naturally occurringas well as synthetic hydroxysterols repress genes containing the sterolresponse element.

It has been suggested that a hydroxycholesterol binding protein servesas a receptor. When the receptor is bound to an oxysterol it acts on thesterol response element to control transcription through a mechanismthat is similar to the action of members of the steroid hormone receptorsuper gene family.

In populations where coronary heart disease is a major health problem,the incidence of the disease is markedly lower in women than in men.This is particularly true in younger age groups, such as men and womenbetween 35 and 44 years of age.

Generally, plasma lipoprotein metabolism is influenced by thecirculating concentrations of gonadal steroids. Changes in serumestrogen and androgen concentrations, resulting from alterations ingonadal status or from the administration of exogenous gonadal steroidsare associated with changes in serum lipoprotein levels. The changeseffected by estrogens and androgens generally support the propositionthat sex differences in lipoproteins are due to hormonal differencesbetween men and women.

The generally accepted relationship between gonadal steroids and plasmalipoproteins is that androgens lower HDL concentrations and increaseLDL, thus contributing to the low HDL and high LDL levels observed inmen when compared to women. Estrogens are held to have opposite effectson lipoproteins; that is HDL is raised and LDL is lowered. These sexsteroid-induced differences in lipoprotein concentrations are thought tocontribute to the lower incidence of cardiovascular disease in womencompared to men. After the menopause, the protective effect of estrogensin women is lost and the incidence of cardiovascular disease increasestowards the male levels. Postmenopausal women who take estrogensgenerally have lower rates of cardiovascular disease than women of asimilar age who do not. Estrogen, particularly when taken orally, lowersplasma levels of LDL and raises those of HDL.

The mechanisms by which estrogen lowers levels of LDL and raises thoseof HDL are not known. In general, changes in the plasma concentration ofa lipoprotein result from changes in the rate of its synthesis or therate of its catabolism. For example, estrogen may lower LDL levels byincreasing the clearance of LDL from plasma, since estrogen increasesthe number of hepatic LDL receptors in animals.

Although estrogens have beneficial effects on serum LDL, given even atvery low levels, long-term estrogen therapy has been implicated in avariety of disorders, including an increase in the risk of uterinecancer and possibly breast cancer, causing many women to avoid thistreatment. Recently suggested therapeutic regimens, which seek to lessenthe cancer risk, such as administering combinations of progestogen andestrogen, cause the patient to experience regular bleeding, which isunacceptable to most older women. Furthermore, combining progesteronewith estrogen seems to blunt the serum cholesterol lowering effects ofestrogen. Concerns over the significant undesirable effects associatedwith estrogen therapy, support the need to develop alternative therapiesfor hypercholesterolemia that generates the desirable effects on serumLDL but does not cause undesirable effects.

Attempts to fill this need by the use of compounds commonly known asantiestrogens, which interact with the estrogen receptor and/or bindwhat has been termed the antiestrogen binding site (AEBS), have hadlimited success, perhaps due to the fact that these compounds generallydisplay a mixed agonist/antagonist effect. That is, although thesecompounds can antagonize estrogen interaction with the receptor, thecompounds themselves may cause estrogenic responses in those tissueshaving estrogen receptors such as the uterus. Therefore someantiestrogens, such as tamoxifen, are subject to some of the sameadverse effects associated with estrogen therapy.

SUMMARY OF THE INVENTION

The invention provides a method of inhibiting bone resorption and boneloss comprising administering to a human in need thereof a compound ofthe formula ##STR2## and pharmaceutically acceptable salts and solvatesthereof, in an amount of about 50 to about 150 mg/day.

The invention also encompasses a method for lowering serum cholesterolcomprising administering a compound of formula I in an amount of about50 to about 150 mg/day.

The invention also encompases pharmaceutical formulations in dosage unitform, comprising, per dosage with an amount of about 50 to about 150 mgof a compound of formula I.

DESCRIPTION OF THE INVENTION

The current invention concerns the discovery that compounds of formula Iare useful for lowering serum cholesterol levels and inhibiting boneresorption and bone loss at dosages of about 50to about 150 mg/day. Themethods provided by this invention are practiced by administering to ahuman in need thereof a dose of a compound of formula I or apharmaceutically acceptable salt or solvate thereof in the amount ofabout 50 to about 150 mg/day, to lower serum cholesterol level, orinhibit bone loss or resorption.

The term "inhibit" is defined to include its generally accepted meaningwhich includes preventing, prohibiting, restraining, and slowing,stopping or reversing progression, or severity, and holding in checkand/or treating existing characteristics. The present method includesboth medical therapeutic and/or prophylactic treatment, as appropriate.

Generally, the compound is formulated with common excipients, diluentsor carriers, and compressed into tablets, or formulated as elixirs orsolutions for convenient oral administration, or administered by theintramuscular or intravenous routes. The compounds can be administeredtransdermally, and are well suited to formulation as sustained releasedosage forms and the like.

The method of the present invention is useful in men as well as women.The substantial absence of estrogenic response should allow men toemploy the method of the present invention without evidencing thefeminizing response of estrogen or estrogen agonists such asgynecomastia. Preferably, however, the methods of the present inventionare useful in women, more preferably estrogen deficient women.

The 2-phenyl-3-aroylbenzothiophene compounds that are the activecomponent in the methods of this invention were first developed by C.David Jones and Tulio Suarez as anti-fertility agents (U.S. Pat. No.4,133,814, issued Jan. 9, 1979). Certain compounds in the group werefound to be useful in suppressing the growth of mammary tumors.

Jones later found a group of related compounds to be useful forantiestrogen and antiandrogen therapy, especially in the treatment ofmammary and prostatic tumors (U.S. Pat. No. 4,418,068, issued Nov. 29,1983). One of these compounds, the hydrochloride salt form of thecompound of formula I, was clinically tested for a brief time for thetreatment of breast cancer. That compound is called raloxifene, formerlykeoxifene.

Raloxifene is currently undergoing human clinical trials for use inosteoporsis and lipid lowering. Draper et al. ("Effects of Raloxifene onBiochemical Markers of Bone and Lipid Metabolism in HealthyPost-Menopausal Women," Fourth International Symposium on Osteoporosis,Hong Kong, Mar. 29, 1993) discussed certain positive findings ofraloxifene's usefulness in inhibiting bone resorption and lowering serumcholesterol. The dosages tested were 200 mg/day and 600 mg/day. Asevidenced by EPO Publication EP-A-584952, published Mar. 2, 1994,(corresponding to U.S. application Ser. No. 07/920,933 filed Jul. 28,1992 (docket X-7947)), the preferred range is listed as 200 mg to 600mg/day. While this dosing range of 200-600 mg/day does providesufficient response and is pharmaceutically acceptable it has now beenfound that a lower dosage range of raloxifene of about 50 mg/day toabout 150 mg/day surprisingly results in providing equivalent benefitsas compared to the higher range.

Raloxifene has been shown to bind to the estrogen receptor and wasoriginally thought to be a molecule whose function and pharmacology wasthat of an anti-estrogen in that it blocked the ability of estrogen toactivate uterine tissue and estrogen dependent breast cancers. Indeed,raloxifene does block the action of estrogen in some cells; however inother cell types, raloxifene activates the same genes as estrogen doesand displays the same pharmacology, e.g., osteoporosis, hyperlipidemia.The unique profile which raloxifene displays and differs from that ofestrogen is now thought to be due to the unique activation and/orsuppression of various gene functions by the raloxifene-estrogenreceptor complex as opposed to the activation and/or suppression ofgenes by the estrogen-estrogen receptor complex. Therefore, althoughraloxifene and estrogen utilize and compete for the same receptor, thepharmacological outcome from gene regulation of the two is not easilypredicted and is unique to each.

Generally, the compound is formulated with common excipients, diluentsor carriers, and compressed into tablets, or formulated as elixirs orsolutions for convenient oral administration, or administered by theintramuscular or intravenous routes. The compounds can be administeredtransdermally or intravaginaly, and may be formulated as sustainedrelease dosage forms and the like.

The compounds used in the methods of the current invention can be madeaccording to established procedures, such as those detailed in U.S. Pat.Nos. 4,133,814, 4,418,068, and 4,380,635 all of which are incorporatedby reference herein. In general, the process starts with abenzo[b]thiophene having a 6-hydroxyl group and a 2-(4-hydroxyphenyl)group. The hydroxyl groups of the starting compound are protected, thethree position is acylated, and the product deprotected to form theformula I compounds. Examples of the preparation of such compounds areprovided in the U.S. patents discussed above.

The compounds used in the methods of this invention formpharmaceutically acceptable acid and base addition salts with a widevariety of organic and inorganic acids and bases and include thephysiologically acceptable salts which are often used in pharmaceuticalchemistry. Such salts are also part of this invention. Typical inorganicacids used to form such salts include hydrochloric, hydrobromic,hydroiodic, nitric, sulfuric, phosphoric, hypophosphoric and the like.Salts derived from organic acids, such as aliphatic mono anddicarboxylic acids, phenyl substituted alkanoic acids, hydroxyalkanoicand hydroxyalkandioic acids, aromatic acids, aliphatic and aromaticsulfonic acids, may also be used. Such pharmaceutically acceptable saltsthus include acetate, phenylacetate, trifluoroacetate, acrylate,ascorbate, benzoate, chlorobenzoate, dinitrobenzoate, hydroxybenzoate,methoxybenzoate, methylbenzoate, o-acetoxybenzoate,naphthalene-2-benzoate, bromide, isobutyrate, phenylbutyrate,β-hydroxybutyrate, butyne-1,4-dioate, hexyne-1,4-dioate, caprate,caprylate, chloride, cinnamate, citrate, formate, fumarate, glycollate,heptanoate, hippurate, lactate, malate, maleate, hydroxymaleate,malonate, mandelate, mesylate, nicotinate, isonicotinate, nitrate,oxalate, phthalate, teraphthalate, phosphate, monohydrogenphosphate,dihydrogenphosphate, metaphosphate, pyrophosphate, propiolate,propionate, phenylpropionate, salicylate, sebacate, succinate, suberate,sulfate, bisulfate, pyrosulfate, sulfite, bisulfite, sulfonate,benzene-sulfonate, p-bromobenzenesulfonate, chlorobenzenesulfonate,ethanesulfonate, 2-hydroxyethanesulfonate, methanesulfonate,naphthalene-1-sulfonate, naphthalene-2-sulfonate, p-toluenesulfonate,xylenesulfonate, tartarate, and the like. A preferred salt is thehydrochloride salt.

The pharmaceutically acceptable acid addition salts are typically formedby reacting a compound of formula I with an equimolar or excess amountof acid. The reactants are generally combined in a mutual solvent suchas diethyl ether or benzene. The salt normally precipitates out ofsolution within about one hour to 10 days and can be isolated byfiltration or the solvent can be stripped off by conventional means.

Bases commonly used for formation of salts include ammonium hydroxideand alkali and alkaline earth metal hydroxides, carbonates, as well asaliphatic and primary, secondary and tertiary amines, aliphaticdiamines. Bases especially useful in the preparation of addition saltsinclude sodium hydroxide, potassium hydroxide, ammonium hydroxide,potassium carbonate, methylamine, diethylamine, ethylene diamine andcyclohexylamine.

The pharmaceutically acceptable salts generally have enhanced solubilitycharacteristics compared to the compound from which they are derived,and thus are often more amenable to formulation as liquids or emulsions.

Pharmaceutical formulations can be prepared by procedures known in theart. For example, the compounds can be formulated with commonexcipients, diluents, or carriers, and formed into tablets, capsules,suspensions, powders, and the like. Examples of excipients, diluents,and carriers that are suitable for such formulations include thefollowing: fillers and extenders such as starch, sugars, mannitol, andsilicic derivatives; binding agents such as carboxymethyl cellulose andother cellulose derivatives, alginates, gelatin, and polyvinylpyrrolidone; moisturizing agents such as glycerol; disintegrating agentssuch as calcium carbonate and sodium bicarbonate; agents for retardingdissolution such as paraffin; resorption accelerators such as quaternaryammonium compounds; surface active agents such as cetyl alcohol,glycerol monostearate; adsorptive carriers such as kaolin and bentonite;and lubricants such as talc, calcium and magnesium stearate, and solidpolyethyl glycols.

The compounds can also be formulated as elixirs or solutions forconvenient oral administration or as solutions appropriate forparenteral administration, for instance by intramuscular, subcutaneousor intravenous routes. Additionally, the compounds are well suited toformulation as sustained release dosage forms and the likes. Theformulations can be so constituted that they release the activeingredient only or preferably in a particular part of the intestinaltract, possibly over a period of time. The coatings, envelopes, andprotective matrices may be made, for example, from polymeric substancesor waxes.

The dosage range for the invention is about 50 to about 150 mg/day, andpreferably 60 to 150 mg/day, and most preferably 60 to 100 mg/day.Particular dosages within the range of the invention were 50, 55, 60,65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115. 120, 125, 130, 135, 140,145, and 150 mg/day.

The compositions are preferably formulated in a unit dosage form eachdosage containing about 50 to about 150 mg, and more preferably theamounts listed above. The term "unit dosage form" refers to physicallydiscrete units, such as tablets and capsules, suitable as unitarydosages, particularly as unitary daily dosages, for human subjects andother mammals, each unit containing a predetermined quantity of activematerial calculated to produce the desired therapeutic effect, inassociation with a suitable pharmaceutical excipient.

The term or period of time of administration to a human subject of thedosage of about 50 to about 150 mg/day will vary depending upon severityof the condition, patient health, and related factors which will bedecided upon by the attending physician. A course of treatment isexpected to be at least for a period of six months, more normally atleast one year, and preferrably on a continual basis.

Examples of formulations using the dosage range follow:

    ______________________________________                                        Formulations                                                                  Ingredient        Quantity (mg/capsule)                                       ______________________________________                                        Formulation 1: Gelatin Capsules                                               Hard gelatin capsules are prepared using the following:                       Raloxifene        50-150                                                      Starch, NF         0-650                                                      Starch flowable powder                                                                           0-650                                                      Silicone fluid 350 centistokes                                                                  0-15                                                        The ingredients are blended, passed through a No. 45 mesh                     U.S. sieve, and filled into hard gelatin capsules.                            Examples of capsule formulations include those shown below:                   Formulation 2: Raloxifene capsule                                             Raloxifene        60                                                          Starch, NF        112                                                         Starch flowable powder                                                                          225.3                                                       Silicone fluid 350 centistokes                                                                  1.7                                                         Formulation 3: Raloxifene capsule                                             Raloxifene        75                                                          Starch, NF        108                                                         Starch flowable powder                                                                          225.3                                                       Silicone fluid 350 centistokes                                                                  1.7                                                         Formulation 4: Raloxifene capsule                                             Raloxifene        100                                                         Starch, NF        103                                                         Starch flowable powder                                                                          225.3                                                       Silicone fluid 350 centistokes                                                                  1.7                                                         Formulation 5: Raloxifene capsule                                             Raloxifene        125                                                         Starch, NF        150                                                         Starch flowable powder                                                                          397                                                         Silicone fluid 350 centistokes                                                                  3.0                                                         Formulation 6: Raloxifene capsule                                             Raloxifene        150                                                         Starch, NF        150                                                         Starch flowable powder                                                                          397                                                         Silicone fluid 350 centistokes                                                                  3.0                                                         ______________________________________                                    

The specific formulations above may be changed in compliance with thereasonable variations provided.

A tablet formulation is prepared using the ingredientsbelow:______________________________________TabletsIngredient Quantity(mg/tablet)______________________________________Formulation7:Raloxifene 60Cellulose, microcrystalline 0-650Silicon dioxide, fumed0-650Stearate acid 0-15Formulation 8:Raloxifene 75Cellulose,microcrystalline 0-650Silicon dioxide, fumed 0-650Stearate acid0-15Formulation 9:Raloxifene 100Cellulose, microcrystalline 0-650Silicondioxide, fumed 0-650Stearate acid 0-15Formulation 10:Raloxifene125Cellulose, microcrystalline 0-650Silicon dioxide, fumed 0-650Stearateacid 0-15Formulation 11:Raloxifene 150Cellulose, microcrystalline0-650Silicon dioxide, fumed 0-650Stearate acid0-15______________________________________

The components are blended and compressed to form tablets.

Alternatively, tablets each containing 50 to 150 mg of active ingredientare made up asfollows:______________________________________TabletsIngredient Quantity(mg/tablet)______________________________________Formulation12:Raloxifene 60Starch 45Cellulose, microcrystalline35Polyvinylpyrrolidone 4(as 10% solution in water)Sodium carboxymethylcellulose 4.5Magnesium stearate 0.5Talc 1Formulation 13:Raloxifene75Starch 45Cellulose, microcrystalline 35Polyvinylpyrrolidone 4(as 10%solution in water)Sodium carboxymethyl cellulose 4.5Magnesium stearate0.5Talc 1Formulation 14:Raloxifene 100Starch 45Cellulose,microcrystalline 35Polyvinylpyrrolidone 4(as 10% solution inwater)Sodium carboxymethyl cellulose 4.5Magnesium stearate 0.5Talc1Formulation 15:Raloxifene 125Starch 45Cellulose, microcrystalline35Polyvinylpyrrolidone 4(as 10% solution in water)Sodium carboxymethylcellulose 4.5Magnesium stearate 0.5Talc 1Formulation 16:Raloxifene150Starch 45Cellulose, microcrystalline 35Polyvinylpyrrolidone 4(as 10%solution in water)Sodium carboxymethyl cellulose 4.5Magnesium stearate0.5Talc 1______________________________________

The active ingredient, starch, and cellulose are passed through a No. 45mesh U.S. sieve and mixed thoroughly. The solution ofpolyvinylpyrrolidone is mixed with the resultant powders which are thenpassed through a No. 14 mesh U.S. sieve. The granules so produced aredried at 50°-60° C. and passed through a No. 18 mesh U.S. sieve. Thesodium carboxymethyl starch, magnesium stearate, and talc, previouslypassed through a No. 60 U.S. sieve, are then added to the granuleswhich, after mixing, are compressed on a tablet machine to yieldtablets.

Suspensions each containing 50-150 mg of medicament per 5 mL dose aremade asfollows:______________________________________SuspensionsIngredientQuantity (mg/5 ml)______________________________________Formulation17:Raloxifene 60 mgSodium carboxymethyl cellulose 50 mgSyrup 1.25mgBenzoic acid solution 0.10 mLFlavor q.v.Color q.v.Purified water to 5mLFormulation 18:Raloxifene 75 mgSodium carboxymethyl cellulose 50mgSyrup 1.25 mgBenzoic acid solution 0.10 mLFlavor q.v.Colorq.v.Purified water to 5 mLFormulation 19:Raloxifene 100 mgSodiumcarboxymethyl cellulose 50 mgSyrup 1.25 mgBenzoic acid solution 0.10mLFlavor q.v.Color q.v.Purified water to 5 mLFormulation 20:Raloxifene125 mgSodium carboxymethyl cellulose 50 mgSyrup 1.25 mgBenzoic acidsolution 0.10 mLFlavor q.v.Color q.v.Purified water to 5 mLFormulation21:Raloxifene 150 mgSodium carboxymethyl cellulose 50 mgSyrup 1.25mgBenzoic acid solution 0.10 mLFlavor q.v.Color q.v.Purified water to 5mL______________________________________

The medicament is passed through a No. 45 mesh U.S. sieve and mixed withthe sodium carboxymethyl cellulose and syrup to form a smooth paste. Thebenzoic acid solution, flavor, and color are diluted:with some of thewater and added, with stirring. Sufficient water is then added toproduce the required volume.

The following examples illustrate the preparation of the compounds usedin the invention.

EXAMPLE 16-hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]benzo[b]thiophene

A 4 g. portion of 6-methanesulfonyloxy-2-(4-methanesulfonyloxyphenyl)-3-[4-(2-piperidinoethoxy)-benzoyl]benzo[b]thiophene, hydrochloride, wascombined with 100 ml. of denatured alcohol and 10 ml. of 5 N sodiumhydroxide, and stirred under reflux for 1.5 hours under a nitrogenatmosphere. The reaction mixture was then evaporated to dryness undervacuum, and the residue was dissolved in 200 ml. of water and washedwith 300 ml. of diethyl ether. The water layer was degassed undervacuum, and then nitrogen was bubbled through it to remove all traces ofether. The mixture Was then acidified with 1 N hydrochloric acid, andthen made basic with excess sodium bicarbonate. The precipitate Wascollected by filtration and washed with cold water to obtain 2.4 g. ofcrude product. It was purified on a 2×30 cm. column of silica gel,eluting first with 700 ml. of 55 methanol in chloroform, followed by 1liter of 10% methanol in chloroform. The impurities came off first, andthe product-containing fractions were combined and evaporated undervacuum to obtain 1.78 g. of yellow oil. The oil was dissolved in 6 m. ofacetone, seeded and chilled in a freezer to obtain 1.2 g. of purifiedproduct, m.p. 143°-147° C. The identity of the product was confirmed asfollows:

nmr spectrum (100 mHz in dmso-d₆)δ1.20-1.65(6H, m. N(CH₂ CH₂)₂ CH₂);2.30-2.45 (4H, m, N(CH₂ CH₂)₂ CH₂); 2.60 (2H, t, J=6 Hz, OCH₂ CH₂ N);4.06(2H, t, J=6Hz, OCH₂ CH₂ N); 6.68 (2H, d, J=9H, aromatic o to OH);6.85 (1H, q, J_(H4-H5) =9 Hz, J_(H5-H7) =2 Hz, H5 of benzothiophenering); 6.90(2H, d, J=9 Hz, aromatic o to OCH₂ CH₂ N); 7.18 (2H, d, J=9Hz, aromatic m to OH); 7.25 (1H, d, J=9z, H4 of benzothiophene ring);7.66 (2H, d, J=9 Hz, aromatic o to CO); 9.72(2H, broad s, OH).Ultraviolet spectrum in ethanol; λ_(max) (ε): 290 nm. (34,000). Electronimpact mass Spectrum M_(t) at m/e 473.

EXAMPLE 26-hydroxy-2-(4-hydoxyphenyl)-3[-4-(2-piperidinoethoxy)benzoyl]benzo[b]thiophene

A 3.6 g. portion of 6-methanesulfonyloxy-2-(4-methanesulfonyloxyphenyl)-3-[4-(2-piperidinoethoxy)-benzoyl]benzo[b]thiophene was dissolved in100 ml. of tetrahydrofuran and 40 ml. of methanol, and 10 ml. of 5 Nsodium hydroxide was added. The mixture was stirred for 16 hours atambient temperature, and was then worked up by the procedure of Example1 above to obtain 3.5 g of a yellow solid. The impure product waspurified by column chromatography on silica gel, eluting with a gradientsolvent from 5% methane in chloroform to 30% methanol in chloroform. Theproduct-containing fractions were evaporated to obtain 1.85 g. of oilyproduct, which was recrystallized from acetone to obtain 1.25 g ofpurified product, m.p. 141°-144° C.

EXAMPLE 36-hydroxy-2-(4-hydroxyphenyl)-3[4-(2-piperidinoethoxy)benzoyl]benzo[b]thiophene,hydrochloride

Under a nitrogen blanket, a mixture of 3 g. of 4-(2-piperidinoethoxy)benzoic acid, hydrochloride, 2 drops of dimethylformamide, 2.5 ml. ofthionyl chloride and 40 ml. of chlorobenzene was heated at 70°-75° C.for about one hour. The excess thionyl chloride and 15-20 ml. of solventwere then distilled off. The remaining suspension was cooled to ambienttemperature, and to it were added 100 ml. of dichloromethane, 2.7 g. of6-methoxy-2-(4-methoxyphenyl)benzo[b]thiophene and 10 g. of aluminumchloride. The solution was stirred for about one hour, 7.5 ml. ofethanethiol was added, and the mixture was stirred for 45 minutes more.Then 40 ml. of tetrahydrofuran was added, followed by 15 ml. of 20%hydrochloric acid, with an exotherm to reflux. Fifty ml. of water and 25ml. of saturated aqueous sodium chloride were added. The mixture wasstirred and allowed to cool to ambient temperature. The precipitate wascollected by filtration and washed successively with 30 ml of water, 40ml of 25% aqueous tetrahydrofuran, and 35 ml. of water. The solids werethen dried at 40° C. under vacuum to obtain 5.05 g. of product, whichwas identified by nmr.

δ1.7(6H, m, N(CH₂ CH₂)₂ CH₂); 2.6-3.1 (2H, m, NCH2); 3.5-4.1 (4H, m,NCH₂ ); 4.4 (2H, m, OCH₂); 6.6-7.4 (9H, m, aromatic); 7.7 (2H, d,aromatic o to CO); 9.8(2H, m, OH).

Test Results

An 8-week parallel, double-blind, placebo study in approximately 160healthy post-menopausal women was completed. The doses of raloxifeneused in this study were 10, 50, and 200 mg. Thee 10 mg dose had nosignificant activity with either bone marker. (See Table I) Because ofdevelopment over time seen with many bone markers, a raloxifene dose of50 mg/day will likely be fully active when evaluated during a study oflonger duration.

                  TABLE I                                                         ______________________________________                                        Baseline Values and Mean (±SEM) Group Changes from                         Baseline to Endpoint in Markers of Bone Metabolism (GGGC)                                      Raloxifene Raloxifene                                                                            Raloxifene                                        Placebo  10 mg      50 mg   200 mg                                    Marker  (n = 42) (n = 42)   (n = 42)                                                                              (n = 41)                                  ______________________________________                                        Serum alkaline phosphatase (U/L)                                              Baseline                                                                              77.31    78.71      73.86   79.07                                             (±3.53)                                                                             (±3.12) (±2.69)                                                                            (±2.77)                                Change  -1.10    0.21       -4.78   -5.93*                                            (±2.08)                                                                             (±1.56) (±1.52)                                                                            (±1.48)                                Serum osteocalcin (ng/mL)                                                     Baseline                                                                              3.94     3.86       3.65    4.21                                              (±0.21)                                                                             (±0.19) (±0.21)                                                                            (±0.21)                                Change  -0.63    -0.27      -0.81   -1.21*                                            (±0.16)                                                                             (±0.13) (±0.15)                                                                            (±0.18)                                ______________________________________                                         Abbreviations: n = greatest number of subjects tested for any one marker;     SEM = standard error of the mean.                                             *Statistically significantly (p < 0.051) different from placebo (twotaile     comparison).                                                             

Serum lipid levels were affected by raloxifene doses of 50 and 200 mg(Table II). Decreases in LDL cholesterol were observed inraloxifene,treated subjects at 50 mg with a comparable decrease in the200 mg patients. Raloxifene-treated subjects showed no changes in HDLlevels. Statistically significant decreases in HDL:LDL ratios and thetotal serum cholesterol levels were observed in raloxifene-treatedsubjects at both the 50 and 200 mg doses.

                  TABLE II                                                        ______________________________________                                        Baseline Values and Mean (±SEM) Group Changes from                         Baseline to Endpoint in Serum Lipids (GGGC)                                                    Raloxifene Raloxifene                                                                            Raloxifene                                        Placebo  10 mg      50 mg   200 mg                                    Variable                                                                              (n = 42) (n = 42)   (n = 42)                                                                              (n = 41)                                  ______________________________________                                        LDL-C (mmol/L)                                                                Baseline                                                                              3.67     4.11#      3.55    3.68                                              (±0.11)                                                                             (±0.17) (±0.16)                                                                            (±0.13)                                Change  0.02     0.05       -0.23*  -0.17                                             (±0.08)                                                                             (±0.10) (±0.06)                                                                            (±0.07)                                HDL-C (mmol/L)                                                                Baseline                                                                              1.41     1.41       1.35    1.32                                              (±0.06)                                                                             (±0.06) (±0.05)                                                                            (±0.05)                                Change  -0.03    0.01       0.04    0.02                                              (±0.03)                                                                             (±0.02) (±0.02)                                                                            (±0.02)                                HDL-C:LDL-C ratio                                                             Baseline                                                                              0.40     0.37       0.42    0.38                                              (±0.02)                                                                             (±0.03) (±0.03)                                                                            (±0.02)                                Change  -0.01    0.00       0.03*   0.03*                                             (±0.01)                                                                             (±0.01) (±0.01)                                                                            (±0.01)                                Toal cholesterol (mmol/L)                                                     Baseline                                                                              5.69     6.18#      5.82    5.71                                              (±0.12)                                                                             (±0.19) (±0.21)                                                                            (±0.14)                                Change  0.10     0.01       -0.23*  -0.15                                             (±0.09)                                                                             (±0.10) (±0.08)                                                                            (±0.08)                                ______________________________________                                         Abbreviations: LDLC = lowdensity lipoprotein cholesterol; HDLC =              highdensity lipoprotein cholesterol; n = greatest number of subjects          tested for any one marker; SEM = standard error of the mean.                  #Statistically significantly (p < 0.050) larger than all other treatments     (twotailed comparison).                                                       *Statistically significantly (p < 0.050) different from placebo (twotaile     comparison).                                                             

I claim:
 1. A method of inhibiting bone loss or bone resorptioncomprising administering to a human in need thereof a compound offormula I ##STR3## or a pharmaceutically acceptable salt or solvatethereof, in an amount of about 50 to 150 mg/day.
 2. A method of claim 1wherein the human has been diagnosed as suffering from osteoporosis. 3.A method of claim 1 wherein the human is a post-menopausal female.
 4. Amethod of claim 1 wherein the human is a male.
 5. A method of claim 1wherein the compound is administered prophylactically.
 6. A method ofclaim 1 wherein the compound of formula I is administered in an amountof about 60 to about 150 mg/day.
 7. A method of claim 1 wherein thecompound of formula I is administered in an amount of 60 mg/day.
 8. Amethod of claim 1 wherein the compound of formula I is administered inan amount of 75 mg/day.
 9. A method of claim 1 wherein the compound offormula I is administered in an amount of 100 mg/day.
 10. A method ofclaim 1 wherein the compound of formula I is administered in an amountof 125 mg/day.
 11. A method of claim 1 wherein the compound of formula Iis administered in an amount of 150 mg/day.
 12. A method of claim 1wherein the compound is the hydrochloride salt.